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Expression and Purification of a Human Pluripotent Stem Cell-Specific Lectin Probe, rBC2LCN

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Lectin Purification and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2132))

Abstract

rBC2LCN is a recombinant N-terminal domain of BC2L-C lectin that is derived from the gram-negative bacteria Burkholderia cenocepacia and specifically binds to Fucα1-2Galβ1-3GlcNAc/GalNAc. Glycome analysis using a high-density lectin microarray revealed that rBC2LCN specifically binds to human pluripotent stem cells (hPSCs) but not to non-hPSCs. The lectin can be added to the cell culture medium for the live staining of hPSCs without causing visible cytotoxicity. Moreover, it can be used in flow cytometric analysis and for the staining of fixed hPSCs. rBC2LCN is a single-chain molecule with a low molecular weight (trimer of 16 kDa), which can be produced in large quantities in Escherichia coli (0.1 g/L). Therefore, rBC2LCN may be a cost-effective probe for use in hPSCs, unlike other hPSC surface marker antibodies that require a mammalian cell expression system for production. In this study, we describe the protocols for the expression and purification of rBC2LCN from E. coli and live staining of hPSCs using this probe.

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Acknowledgments

We thank the RIKEN BioResource Center for providing the hiPS cell line 201B7 (HPS0063).

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Authors and Affiliations

  1. Biotechnology Research Institute for Drug Discovery (BRD), National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan

    Hiroaki Tateno

  2. Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance, University of Tsukuba, Tsukuba, Japan

    Hiroaki Tateno

  3. JST PRESTO, Tsukuba, Ibaraki, Japan

    Hiroaki Tateno

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  1. Hiroaki Tateno
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Correspondence to Hiroaki Tateno .

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Editors and Affiliations

  1. Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan

    Jun Hirabayashi

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Tateno, H. (2020). Expression and Purification of a Human Pluripotent Stem Cell-Specific Lectin Probe, rBC2LCN. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_44

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  • DOI: https://doi.org/10.1007/978-1-0716-0430-4_44

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0429-8

  • Online ISBN: 978-1-0716-0430-4

  • eBook Packages: Springer Protocols

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